TLRs 2 and 4 are not involved in hypersusceptibility to acute Pseudomonas aeruginosa lung infections

TLRs are implicated in defense against microorganisms. Animal models have demonstrated that the susceptibility to a number of Gram-negative pathogens is linked to TLR4, and thus LPS of many Gram-negative bacteria have been implicated as virulence factors. To assess the role of this pathogen-associated molecular pattern as it is exposed on intact Pseudomonas aeruginosa, the susceptibility of mice lacking TLR4 or both TLR2 and TLR4 was examined in a model of acute Pseudomonas pneumonia. These mutant mice were not hypersusceptible to the Pseudomonas challenge and mounted an effective innate response that cleared the organism despite low levels of TNF-alpha and KC in the airways. Bacterial and neutrophil counts in the lung were similar in control and TLR-deficient mice at 6 and 24 h after infection. MyD88(-/-) mice were, however, hypersusceptible, with 100% of mice dying within 48 h with a lower dose of P. aeruginosa. Of note there were normal levels of IL-6 and G-CSF in the airways of TLR mutant mice that were absent from the MyD88(-/-) mice. Thus, the susceptibility of mice to P. aeruginosa acute lung infection does not go through TLR2 or TLR4, implying that Pseudomonas LPS is not the most important virulence factor in acute pneumonia caused by this organism. Furthermore, G-CSF treatment of infected MyD88(-/-) mice results in improved clearance and survival. Thus, the resistance to infection in TLR2/TLR4(-/-) mice may be linked to G-CSF and possibly IL-6 production.     

Phenotypic variation in adult behavioral response and offspring fitness in Plutella xylostella (Lepidoptera: Plutellidae) in response to permethrin

Phenotypic variation in behavioral response and physiological tolerance to permethrin was measured and compared between two populations of diamondback moth, Plutella xylostella (L.). A field population from Celeryville, OH, and a laboratory population originally collected from Wooster, OH, were compared. In laboratory choice bioassays, females from both populations were less likely to oviposit on cabbage leaf disks and seedlings treated with permethrin. The oviposition-deterrent effect was positively related to permethrin concentration. The laboratory population was significantly more behaviorally responsive to the insecticide and showed a significantly greater avoidance than the field population of the highest concentration of permethrin tested (1.50 g [AI] /liter). The physiological response of each population was measured by feeding bioassays, and the laboratory population was physiologically more susceptible to the permethrin. Larvae from the laboratory population that were fed permethrin-treated leaves had significantly lower growth rate, higher mortality, and lower adult fecundity compared with larvae from the field population. These data suggest that female moths that are more behaviorally responsive to permethrin produce offspring that tend to be more susceptible to the same insecticide, at least for the populations tested. The relationship between adult behavioral response and larval physiological tolerance to permethrin is discussed.     

Efficient transduction and selection of human T-lymphocytes with bicistronic Thy1/HSV1-TK retroviral vector produced by a human packaging cell line

BACKGROUND: T-cells expressing the HSV1-TK suicide gene can be used for the control of graft-versus-host disease following allogeneic stem cell transplantation. To develop clinical trials based on such a strategy, we have generated under good manufacturing procedures a novel 'split genome' human packaging cell line (1704 cells). METHODS: To minimize the risk of generating replication-competent retroviruses, pol was truncated to remove sequences overlapping with env. To improve retroviral infection and selection of transduced T-cells, high titers of GALV-pseudotyped retroviral particles harboring a bicistronic Thy1-IRES-TK vector coding for the CD90 GPI-anchored membrane molecule were produced by 1704 cells. RESULTS: Using 1704 cell supernatant and an optimized transduction protocol, approximately 50% of primary T-cells were transduced and could then be purified (approximately 95%) using clinical-grade immunomagnetic beads directed against CD90. Over 96% of these OKT3/IL-2-activated CD90(+)-selected T-cells were killed by ganciclovir. Cell proliferation and cytokine production of transduced T-cells and HLA-restricted cytotoxicity of transduced T-cell clones were identical to those of their non-transduced counterparts cultured under the same conditions. CONCLUSIONS: GALV-pseudotyped retroviral particles harboring a bicistronic Thy1-IRES-TK vector allow efficient transduction and rapid selection of human T-cells under conditions applicable for clinical trials using the new human 1704 packaging cell line.     

Immobilized metal-ion affinity chromatography of human antibodies and their proteolytic fragments

Immobilized metal-ion affinity chromatography (IMAC) performed with four different transition metal ions: copper(II), nickel(II), zinc(II) and cobalt(II), was used to study the adsorption properties of human polyclonal gamma-globulines (IgG), Cohn II-III fractions, and their pepsin cleaved fragments: Fab'2 and F'c. In each case, digested products showed lower affinity for metal ions, as well by decreasing pH elution as by competition with imidazole. An explanation was proposed by the presence of a histidine (His) cluster in the F'c domain of IgGs, identified by computer calculation (accessible surface area (ASA) determination) as the more probable His 433-x-His 435 sequence presented in the CH3 domain of human IgG heavy chain. As shown by IMAC and electrophoresis, F'c and undigested IgG have higher affinity for transition metal ions than Fab'2 fragments and could be then separated in one step by IMAC. When chelated Zn(II) or Co(II) are used as ligands, the Fab'2 fragment could be easily recovered under mild conditions (pH 7) in the non-retained fraction. This approach could be used as a powerful alternative to conventional protein A/G methods for the commercial preparation of non immunogen active Fab'2 fragments.     

Hydrous and photosynthetic adaptations of common and durum wheat to saline stress

Seven varieties of bred wheat and seven varieties of durum wheat were cultivated in three different sites from the area of Errachidia (southeastern Morocco). These sites differ by the degree of salinity in the irrigation water. Results obtained showed that the reduction in leaf area is the principal strategy that makes it possible to attenuate the effects of the reduction in the availability of water under saline stress. Bread wheat, which limited the reduction in the leaf area, with the risk to undergo some hydrous problems, seems to better preserve its photosynthetic potentialities and grain productivity.     

The Introduced Green Alga Caulerpa Taxifolia Continues to Spread in the Mediterranean

The tropical green alga Caulerpa taxifolia in the Mediterranean has spread steadily since its introduction in 1984. At the end of 2000, approximately 131km² of benthos had been colonized in 103 independent areas along 191km of coastline in six countries (Spain, France, Monaco, Italy, Croatia and Tunisia). Large regions neighboring the invaded areas appear favorable to further colonization, and there is thus no reason to believe that spreading will slow down in the years to come.     

Découverte de charophytes et ostracodes de l’Yprésien inférieur dans les Monts des Ksour (Algérie) : biostratigraphie et paléoécologie

A stratigraphic, palaeontological and sedimentological study was carried out in the red beds cropping out on the left side of Oued Tafarahit, south-east of the Ksour Mountains (Algeria). The studied succession consists of a non-marine fining-upward detrital formation, including microconglomerate, sandstone and clay beds, which were previously attributed to the Cenozoic sensu lato, since no palaeontological evidence was available. Sedimentology suggests deposition in a fluvial environment. The clayey levels yielded the assemblage formed by the charophytes Peckichara atlasensis, Maedleriella cristellata, Nitellopsis (Tectochara) thaleri, Grovesichara sp., and Lamprothamnium papulosum and the ostracods Neocyprideis meguerchiensis, Herpetocypris? sp. and Cyprinotus? sp. This assemblage allows constraining the age of the detrital series from Oued Tafarahit to the Ypresian. Ostracods are typical of freshwater to euryhaline environments. The freshwater taxa (Herpetocypris? sp. and Cyprinotus? sp.) would indicate phases of desalinisation during periods of flood. However, the brackish water species (Neocyprideis meguerchiensis) is characteristic of saline phases related to low water table periods. Moreover the charophyte and ostracod assemblage confirms a close palaeobiogeographic relationship between North Africa and Southern Europe during the Lower Eocene.     

A Fish Assemblage from the Middle Eocene from Libya (Dur At-Talah) and the Earliest Record of Modern African Fish Genera

In the early nineteen sixties, Arambourg and Magnier found some freshwater fish (i.e., Polypterus sp., Siluriformes indet. and Lates sp.) mixed with marine members in an Eocene vertebrate assemblage at Gebel Coquin, in the southern Libyan Desert. This locality, aged ca 37–39Ma and now known under the name of Dur At-Talah, has been recently excavated. A new fish assemblage, mostly composed of teeth, was collected by the Mission Paléontologique Franco-Libyenne. In this paper, we describe freshwater fish members including a dipnoan (Protopterus sp.), and several actinopterygians: bichir (Polypterus sp.), aba fish (Gymnarchus sp.), several catfishes (Chrysichthys sp. and a mochokid indet.), several characiforms (including the tiger fish Hydrocynus sp., and one or two alestin-like fish), and perciforms (including the snake-head fish Parachanna sp. and at least one cichlid). Together with the fossiliferous outcrops at Birket Qarun in Egypt, the Libyan site at Dur At-Talah reduces a 10-Ma chronological gap in the fossil record of African freshwater fish. Their fish assemblages overlap in their composition and thus constitute a rather homogenous, original and significant amount of new elements regarding the Paleogene African ichthyofauna. This supports the establishment of the modern African freshwater fish fauna during this time period because these sites mostly contain the earliest members known in modern genera.     

Genetic diversity among summer and winter Beauveria bassiana populations as revealed by AFLP analysis

The genetic diversity of Beauveria bassiana was investigated by comparing 40 isolates collected from summer and overwintering populations of Sunn pest from different areas in Syria and Turkey, using amplified fragment length polymorphism (AFLP) markers. Considerable genetic variability among B. bassiana isolates was revealed. The examined isolates were divided into three distinct clusters (A, B, and C). Within these clusters, the summer isolates from Syria and Turkey were grouped together in three sub-clusters (A3, A4, and B2). Also, principal coordinate analyses (PCA) showed clear separation (62.5%) between summer and winter isolates. These differences in the genetic structure may be explained by the variety of eco-geography over the sampled areas of B. bassiana isolates. This information on genetic variation among summer and winter B. bassiana isolates is helpful in designing an effective integrated pest management program for Sunn pest.